一步净化结合超高液相色谱串联质谱同时测定雅鱼中13类54种兽药残留  

One-step Purification Combined with Ultra-High-Performance Liquid Chromatography-Tandem Mass Spectrometry for the Determination of Residues of 54 Veterinary Drugs Belonging to 13 Classes in“Ya-Fish”

作　　者：杨巧慧 刘中良 陈亚 李霞雪 张宇 曾艳 张建雄 闻瑞琪 兰韬[4] YANG Qiaohui;LIU Zhongliang;CHEN Ya;LI Xiaxue;ZHANG Yu;ZENG Yan;ZHANG Jianxiong;WEN Ruiqi;LAN Tao(Ya'an Agricultural Product Quality Monitoring and Inspection Center,Ya'an 625000,China;Tangshan Food and Drug Comprehensive Inspection and Test Center,Tangshan 063000,China;Beijing KNORTH Technology Co.,Ltd.,Beijing 102299,China;China National Institute of Standardization,Beijing 100191,China)

机构地区：[1]雅安市农产品质量监测检验中心,四川雅安625000 [2]唐山市食品药品综合检验检测中心,河北唐山063000 [3]北京科德诺思技术有限公司,北京102299 [4]中国标准化研究院,北京100191

出　　处：《食品工业科技》2024年第3期242-252,共11页Science and Technology of Food Industry

基　　金：四川省农产品质量安全风险监测项目(川农函[2022]115号);中央级公益性科研院所基本科研业务费(562022Y-9418);国家市场监督管理总局技术保障专项项目(2021MK159)。

摘　　要：目的:本研究建立了一种一步净化法结合液相色谱串联质谱(LC-MS/MS)同时检测雅鱼(Schizothorax prenanti)中13类54种兽药残留的快速检测方法。方法:向2.0 g样品中加入8 mL 0.2%甲酸-乙腈:水溶液(90:10,V/V)提取,冰水浴超声提取,LPAS一步净化。采用C18柱,甲醇-0.1%甲酸/5 mmol/L乙酸铵溶液梯度洗脱,动态多反应监测(dMRM),正/负离子模式,内标法定量,LC-MS/MS测定。以添加54种兽药的方式,对提取方式、提取体积、盐的加入、液相和质谱条件进行了优化,对方法性能(基质效应、方法检出限、方法定量限、准确度、精密度)等进行了验证。结果:54种兽药在0.1~50μg/L范围内线性关系良好,决定系数(R^(2))≥0.992,方法检出限(LOD)为0.03~2.5μg/kg之间,方法定量限(LOQ)为0.1~8.3μg/kg之间,在低(添加量5~10μg/kg)、中(添加量10~20μg/kg)、高(添加量50μg/kg)三个水平下54种药物的平均回收率为70.45%~118.1%,RSD为0.01%~9.9%。对50份养殖雅鱼样本应用该方法进行检测,7份样品有药物检出,恩诺沙星和达氟沙星各检出2次,磺胺甲噁唑检出4次,均低于定量限的定性检出。结论:该方法操作简单,一步净化,且方法检出限低、灵敏度高,准确度和精密度均较好,方法性能验证其符合多种兽药残留要求,可为大批量检测雅鱼中多种类兽药残留提供有力支持。Objective:In this study,a one-step purification method combined with liquid chromatography tandem mass spectrometry(LC-MS/MS)was established for the simultaneous detection of 54 veterinary drug residues belongs to 13 classes of ya-fish(Schizothorax prenati).Methods:8 mL of 0.2%formic acid-acetonitrile:water solution(90:10,V/V)was added to 2.0 g sample and extracted by ultrasonication in an ice-water bath and cleaned up in one step by LPAS.The determination was performed on a C18 column with a gradient elution of methanol-0.1%formic acid-5 mmol/L ammonium acetate solution,dynamic multiple reaction monitoring(dMRM),positive/negative ionization mode,and quantitative LC-MS/MS by internal standard method.By adding 54 veterinary drugs,the extraction method,extraction volume,salt addition,liquid phase and mass spectrometry conditions were optimized,and the method performance(matrix effect,detection limit,quantification limit,accuracy and precision)were verified.Results:The linear relationships between the 54 drugs and their concentrations were good within the range of 0.1~50μg/L,coefficient of determination(R^(2))≥0.992.The limits of detection(LODs)were between 0.03~2.5μg/kg and the limits of quantification(LOQs)were 0.1~8.3μg/kg.The average recoveries of the 54 drugs at low(5~10μg/kg),medium(10~20μg/kg)and the high level(50μg/kg)ranged from 70.45%to 118.1%,with relative standard deviations(RSD)between 0.01%and 9.9%.This method was applied to detect the drug in 50 samples of cultured ya-fish,and drugs were detected in seven of them.Enrofloxacin and danofloxacin were each detected twice,and sulfamethoxazole was detected four times,all at levels below the limit of quantification.Conclusion:The method was simple to operate,one-step purification,and had low limits,high sensitivity,good accuracy and precision,which met the technical requirements of multi-veterinary drug residue detection,and can provide technical support for the rapid and efficient detection of various veterinary drug residues in large quantities of

关 键 词：一步净化 液相色谱串联质谱 雅鱼 脂肪蛋白吸附 兽药残留 

分 类 号：TS254.7[轻工技术与工程—水产品加工及贮藏工程]